DE1518349B2 - Process for the preparation of eledoisin-active compounds - Google Patents

Description

R-L-Alanyl-L-phenylalanyl-L-a-aminoacylglycyl-L-leucyl-L-methionine amide,

in which R denotes formyl, nicotinyl, a-hydroxyisovaleryl, Chloroacetyl-, a-acetyl-L-lysyl-, α, ε-diacetyl-L-lysyl- or ε-aminocapronyl radical and L-α-aminoacyl for the L-isoleucyl or L-valyl radical, characterized in that an acid of the formula ROH is mixed with L-alanine, L-phenylalanine, L-isoleucine or L-valine, glycine, L-leucine and L-methionine amide or their reactive derivatives according to in protein chemistry known methods.

In another additional patent 1518 344 has shown that the radical R can be any natural, non-acylated amino acid without the eledoisin effect the peptides decrease. It has now been found that not only the compounds in which R is a natural, not acylated amino acid residue, have an eledoisin-like effect, but surprisingly in increased Measure those compounds in which R denotes the ίο formyl, nicotinyl, a-hydroxyisovaleryl, chloroacetyl, α-acetyl-L-lysyl, α-ε-diacetyl-L-lysyl or -.- .. ε-aminocapronyl radical.

The invention accordingly relates to compounds of the general formula

R-L-Alanyl-L-phenylalanyl-L-a-aminoacylglycyl-L-leucyl-L-methionine amide,

in which R denotes formyl, nicotinyl, a-hydroxyisovaleryl, chloroacetyl, a-acetyl-L-lysyl, α, ε-diacetyl-L-lysyl or ε-aminocapronyl radical, and L-α-aminoacyl stands for the L-isoleucyl or L-valyl radical. The following table gives an overview of the biological activity of the compounds

R-Ala-Phe-Ileu-Gly-Leu-Met-NHa:

Tabel R. Relative lowering Bradykinin = 1 of blood pressure am anesthetized Formyl Dog related 20th Chloroacetyl on eledoisin = 1 20th L-a-hydroxyiso- about 2 valeryl 2 20th ε-aminocapronyl about 25 Nicotinyl 2 around 15 4 ° a-monoacetyl- about 2.5 lysyi 1.5 35 3.5

45

From French patent 1,329,840 it is known that the heptapeptide

. L-asparagyl-L-alanyl-L-phenylalanyl-L-isoleucylglycyl-L-leucyl-L-methionine amide,

whose free carboxyl group on the asparagyl radical can also be modified by amide formation, a has a strong, especially expanding effect on the blood vessel system.

The main patent 1 518 340 describes the synthesis of peptides of the general formula

R-L-Alanyl-L-phenylalanyl-L-α-aminoacylglycyl-L-leucyl-L-methionine amide

described, in which R stands for a hydrogen atom, the L-asparaginyl or L-glutaminyl and a-aminoacyl stands for the isoleucyl or valyl radical, while the additional patent 1 518 342 teaches that R also denotes Can mean glycyl radical. The compounds of the general formula show one of the eledoisin, one mainly antihypertensive undekapeptide, comparable biological activity.

The greater effect of the compounds according to the invention is unexpected in that they are not can be explained by the prevented enzymatic degradation with aminopeptidases. This proves the the effect of the diacetyl-lysyl derivative and the low activity only correspond to that of the eledoisin of the capronyl and caprinoyl compounds (30% and 10% of the eledoisin, respectively), which are also not with Aminopeptidases are degradable.

In many cases, the compounds according to the invention are also distinguished from those hitherto known products are characterized by their ease of manufacture, for example because they are arbitrarily substituted organic acids are often more accessible than many amino acids. The synthesis of the new compounds takes place according to methods known per se.

Example 1 Formyl-ala-Phe-Ileu-Gly-Leu-Met-NH ₂

Formyl-Ala-Phe-OMe was obtained in the usual manner by the anhydride method [m.p. 115 to 117 ⁰ C (from ethanol / petroleum ether), [tx] _D = -10.8 ° (c = 0.8; DMF)] and converted into the hydrazide with four times the amount of hydrazine hydrate. M.p. 207

ι. u ι. υ jtc?
3 4

to 208 ⁰ C (from DMF / petroleum ether), [<x] _D = - 49.9 ° the usual washing of the solid substance and

(c = 0.5; glacial acetic acid). Falling from DMF / aqueous citric acid was

Conversion of 0.3 g of the hydrazide (azide formation, the yield 2.8 g (61.5%); melting point 249 to 254 ° C,

using tert-butyl nitrite in hydrogen chloride / tetra- [α]? = -40.8 ° (c = 0.5; glacial acetic acid),
hydrofuran) with 0.6 g of H-Ileu-Gly-Leu-Met-NH ₂ · HCl 5

resulted in 0.5 g of the desired compound, mp. 267 c) Ac-L-Lys-L-Ala-L-Phe-L-Ileu-Gly-L-Leu-L-Metbis 268 ⁰ C, [<x] _D = -42.0 ° (c = 0.5; DMF). . NH ₂

Example 2 was in quantitative yield by protecting groups

lo cleavage with HCl / glacial acetic acid from the nicotinoyl-Ala-Phe-Ileu-Gly-Leu-Met-NH ₂ th compound shown in 5 b).

Mp. 225 to 245 ° C, [*]? = -42.5 ° (c = 0.5; ice-cream from 3.2 g nicotinic acid chloride and 5.7 g vinegar).

H-Ala-Phe-OMe · HCl were in chloroform according to Example 6

Addition of 8.4 ecm of triethylamine, 5.6 g of nicotinoyl-15

Ala-Phe-OMe, mp. 161-162 ⁰ C (from a) BOC-L-Lys (Ac) -OH

Ethanol / petroleum ether), [oc] _D = + 32.0 ° (c = 0.5; glacial acetic acid). 2.3 g (12 mmol) of HL-LyS- (Ac) -OH were dissolved in hydrazinolysis with four times the amount of hydrazine-dioxane / water 1: 1 and in the presence of hydrate gave the hydrazide [mp. 209 to 211 ° C, 20 1.5 g (36 mmol) of magnesium oxide with 3.4 g (14.2 mmol) [α] ΰ = 16.6 ° (c = 0.5; glacial acetic acid), the tert as in Example 1 .-Butyloxycarbonyl-p-nitro-phenyl ester reacted, reacted with H-Ileu-Gly-Leu-Met-NH ₂ · HCl, yield: 1.31 g (38%); Mp 137-138 ° C (mp of 272-274 ⁰ C, [<x] _D = -27.5 ° (c = 0.5;. DMF) Essigester / petroleum ether.). [«]? = -10.1 ° (c = 0.5; glacial acetic acid).
Example3 "25

b) BOC-L-LyS- (Ac) -OPhNO ₂
α-Hydroxyisovaleryl-Ala-Phe-Ileu-Gly-Leu-Met-NH ₂

1.0 g (3.4 mmol) BOC-L-LyS- (Ac) -OH was with

From 0.3 g of La-hydroxyisovaleric acid hydrazide, 0.6 g (4.5 mmol) of p-nitrophenol according to the Carbodi was esterified with 0.3 ecm of tert-butyl nitrite in 2.9 ecm of a 30 imide method. Yield: 1.3 g (92%); Mp. 96 1.5 η solution of hydrogen chloride in tetrahydrofuran up to 98 ° C (from ethyl acetate / petroleum ether), [<x] $ = -24.3 ° the azide is produced, which after neutralization (c = 0.5; Ethanol),
with triethylamine with 1.5 g H-Ala-Phe-Ileu-Gly-Leu-

Met-NH ₂ ■ HCl and 0.3 ecm of triethylamine reacted c) BOC-L-Lys- (Ac) -L-Ala-L-Phe-L-Ileu-Gly-

became. Yield 1.3 g, m.p. 267 to -27O ⁰ C, [tx] _D 35 L-Leu-L-Met-NH ₂

= -38.7 (c = 0.5; DMF).

1.2 g (3 mmol) of BOC-L-LyS- (Ac) -OPhNO ₂ were

B e i s ρ i e 1 4 with 2.0 g (3 mmol) H-L-Ala-L-Phe-L-Ileu-Gly-L-Leu-

L-Met-NH ₂ reacted in DMF for 3 days at 40 ° C. To

Chloracetyl-Ala-Phe-Ileu-Gly-Leu-Met-NHsä 40 of the usual work-up were 1.3 g (48%)

obtain. Mp. 239 to 260 ⁰ C, [«]? = -44.0 ° (c = 0.5; 0.2 g of monochloroacetic acid were mixed with 0.3 ecm of glacial acetic acid).
Triethylamine and 0.2 ecm of ethyl chloroformate converted into the mixed anhydride and d) HL-Lys- (Ac) -L-Ala-L-Phe-L-Ileu-Gly-L-Leudieses with 1.4 g of H-Ala-Phe -Ileu-Gly-Leu-Met-NH ₂ 45 L-Met-NH ₂
coupled.

Yield 1.2 g, melting point 238 to 242 ° C., [<x] d = -36.4 °. The protected compound was converted into protection (c = 0.5; DMF). group cleavage obtained with HCl / glacial acetic acid. Fp. About

230 ° C (dec.); [a] f = -27.0 ° (c = 0.5; glacial acetic acid). B eis ρ ie 1 5 50 C ₃₉ H ₆₅ N ₉ O ₈ S 1.6 HCl 3 H ₂ O (932 45)

a) Ac-L-Lys- (BOC) -NHNH, e) Ac-L-Lys- (Ac) -L-Ala-L-Phe-Ileu-

Gly-L-Leu-L-Met-NH ₂

4.0 g (15.4 mmol) of H-L-LyS- (BOC) -OMe were

with 2.8 g (15.4 mmol) of acetic acid p-nitrophenyl ester 55 800 mg (0.9 mmol) of a-acetyl-lysine-heptapeptide-

acetylated. After the release, the methyl ester amide hydrochloride, which was only obtained as an oil, became with

[3.9 g (84.1%); [oc] f = -18.7 ° (c = 1.5; pyridine)] 0.13 ecm triethylamine in DMF with 360 mg (2 mmol)

were treated in the usual way by hydrazinolysis with 2.5 times the amount of p-nitrophenyl acetate

Hydrazine hydrate produced the hydrazide. Yield: sets. Yield: 640 mg (80%), melting point 260 to 266 ° C,

2.1 g (55%), melting point 123 to 125 ° C (from ethyl acetate). 60 [«]? = -44.8 ° (c = 0.5; glacial acetic acid).

b) Ac-L-Lys- (BOC) -L-Ala-L-Phe-L-Ileu-Gly-L-Leu Example 7

L-Met-NH _{2 a} -j _£ .BOC-aminocaproic acid- (BOC-e-Acap-OH)

1.5 g (5 mmol) of Ac-L-LyS- (BOC) -NHNH ₂ were 65 From the ε-aminocaproic acid with tert-butyloxy

with 0.5 ecm (5.2 mmol) of tert-butyl nitrite in the azide carbonyl-p-nitrophenyl ester are in the usual

transferred and this obtained with 3.2 g (5 mmol) of H-L-Ala-way 77% of a crystallizing oil.

L-Phe-L-Ileu-Gly-L-Leu-L-Met-NH ₂ coupled. After m.p. 30 to 31 ° C.

5 6

b) BOC-fi-Acap-L-Alä-L-Phe-L-Ileu-Gly-L-Leu-

..- .. '· .. L ^ Met-NH ₂ c) He-Acap-L-Ala-L-Phe-L-Ileu-Gly-L-Leu-

L-Met-NH ₂ ■ HCl

According to the anhydride method, 1.4 g

(6mMol) BOC-e-aminocaproic acid and 3.9g (6mMol) 5 by splitting off protective groups with HCl / glacial acetic acid

H-L-Ala-L-Phe-L-Ileu-Gly-L-Leu-L-Met-NHa 4.1 g was the hydrochloride in quantitative yield

(78%) received. Mp 247-248 ° C; [«]? = -33.6 ° obtained. Mp. 221 to 230 ⁰ C, [«]? = -35.0 ° (c = 0.5;

(c = 0.6 DMF). DMF).

Claims (9)

Patent claims: 1. Compounds of the general formula R-L-Alanyl-L-phenylalanyl-L-a-aminoacylglycyl-L-leucyl-L-methionine amide, in which R denotes the formyl, nicotinyl, a-hydroxyisovaleryl, chloroacetyl, a-acetyl-L-lysyl, α, ε-diacetyl-L-lysyl or ε-aminocapronyl radical and La-aminoacyl for the L- Is isoleucyl or L-valyl radical. . ^: '· ^: ·' / -, -. ··> 2. Formyl-L-alanyl-L-phenylalanyl-L-isoleucylglycyl-L-leucyl-L-methionine amide. .. ■ 3. Nicotinyl-L-alanyl-L-phenylalanyl-L-isoleucyl-glycyl-L-leucyl-L-methionine amide. 4. α-Hydroxyisovaleryl-L-alanyl-L-phenylalanyl-L-isoleucyl-glycyl-L-leucyl-L-methionine amide. 5. Chloroacetyl-L-alanyl-L-phenylalanyl-L-isoleucyl-glycyl-L-leucyl-L-methionine amide. 6. oi- Acetyl-L- lysyl - L -alanyl -L - phenylalanyl L-isoleucyl-glycyl-L-leucyl-L-methionine amide. 7. α, ε-Diacetyl-L-lysyl-L-alanyl-L-phenylalanyl-L-isoleucyl-glycyl-L-leucyl-L-methionine amide. 8. ε-Aminocapronyl - L - alanyl - L-phenylalanyl-L-isoleucyl-glycyl-L-leucyl-L-methionine amide. 9. Process according to Patent 1,518,340 for the preparation of peptides of the general formula